體外仿生肺肝纖維化3D模型研究;Establishment of a 3D dynamic model of in vitro biomimetic lung and liver fibrosis

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Title:	體外仿生肺肝纖維化3D模型研究;Establishment of a 3D dynamic model of in vitro biomimetic lung and liver fibrosis
Authors:	李奕鋆;Li, Yi-Yun
Contributors:	系統生物與生物資訊研究所
Keywords:	循環拉伸;3D 模型;纖維化;Cyclic Stretch;3D model;fibrosis
Date:	2022-08-20
Issue Date:	2022-10-04 11:15:56 (UTC+8)
Publisher:	國立中央大學
Abstract:	在人體內組織和細胞會受到不同程度外力刺激，在纖維化過程中機械刺激很大的影響纖維化程度，且週期性的拉伸可以調控許多的細胞行為，包括細胞的生長、分化、基因表達和訊息傳遞。另外體內的細胞行為涉及細胞與周遭微環境之間的動態相互作用，因此本實驗使用甲基丙烯酸酐化魚明膠作為三維支架模擬體內的微環境進行機械循環拉伸，拉伸條件為應變量 15%、頻率 0.5 Hz，然後使用商業用矽膠培養膜作為二維培養模型以相同的拉伸條件比較兩種材料對於纖維化程度的影響，實驗的對象分別是人類肝星狀細胞株 LX2 以及人類肺成纖維細胞株 HPF，從免疫螢光染色結果發現三維支架培養的 LX2 和 HPF 在拉伸 24 和 48 小時後肌成纖維標誌物 α-SMA 的表現量提升，在酵素免疫分析、即時定量聚合酶連鎖反應和西方墨點法也驗證了三維仿生動態培養更能夠促進細胞纖維化。;Cells and tissues in the human body are subjected to forces of varying degrees. During fibrosis, mechanical stimulation greatly affects fibrosis, and cyclic stretch can regulate many cellular behaviors, including cell growth, differentiation, gene expression and messaging. In addition, the behavior of cells in vivo involves a dynamic interaction between cells and the surrounding microenvironment. Therefore, in this experiment, fish gelatin methacryloyl was used as a three dimensional scaffold to simulate the microenvironment in vivo under mechanical cyclic stretching. The condition was 15% strain, 0.5 Hz frequency, and then a polydimethylsiloxane membrane was used as a 2D culture model to compare the effect of the two materials on the degree of fibrosis under the same stretch conditions. Human hepatic stellate cell LX2 and human pulmonary fibroblasts HPF were subjected to cyclic stretching. The results show that immunofluorescent staining in LX2 and HPF cultured on three-dimensional scaffolds were expressed more myofibroblast marker α-SMA after stretching for 24 and 48 hours. The results of enzyme-linked immunosorbent assay, real-time quantitative polymerase chain reaction and western blot also confirmed that 3D biomimetic dynamic culture is more conducive to cellular fibrosis.
Appears in Collections:	[Institute of Systems Biology and Bioinformatics] Electronic Thesis & Dissertation

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